Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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    The Synchytrium endobioticum AvrSen1 triggers a Hypersensitive Response in Sen1 potatoes while natural variants evade detection
    Vossenberg, Bart van de; Prodhomme, Charlotte ; Arkel, Gert van; Gent-Pelzer, Marga van; Bergervoet-van Deelen, Marjan ; Brankovics, Balázs ; Przetakiewicz, J. ; Visser, Richard ; Lee, Theo van der; Vossen, Jack - \ 2019
    Wageningen University and Research
    PRJEB31420 - ERP113980 - Synchytrium endobioticum
    Synchytrium endobioticum is an obligate biotrophic fungus of the phylum Chytridiomycota. It causes potato wart disease, has a world-wide quarantine status and is included on the HHS and USDA Select Agent list. S. endobioticum isolates are grouped in pathotypes based on their ability to evade host-resistance in a set of differential potato varieties. So far, thirty-nine pathotypes are reported. A single dominant gene (Sen1) governs pathotype 1 resistance and we anticipated that the underlying molecular model would involve a pathogen effector (AvrSen1) that is recognized by the host.The S. endobioticum specific secretome of fourteen isolates representing six different pathotypes was screened for effectors specifically present in pathotype 1(D1) isolates but absent in others. We discovered a single AvrSen1 candidate. Expression of this candidate in potato Sen1 plants showed a specific hypersensitive response, which co-segregated with the Sen1 resistance in potato populations. No HR was obtained with truncated genes found in pathotypes that evaded recognition by Sen1. These findings established that our candidate gene was indeed Avrsen1. AvrSen1 is a single copy gene and encodes a 376 amino acid protein without predicted function or functional domains, and is the first effector gene identified in Chytridiomycota.
    The Synchytrium endobioticum AvrSen1 triggers a Hypersensitive Response in Sen1 potatoes while natural variants evade detection
    Vossenberg, Bart van de; Prodhomme, Charlotte ; Arkel, G. van; Gent-Pelzer, M.P.E. van; Bergervoet-van Deelen, J.E.M. ; Brankovics, Balázs ; Przetakiewicz, J. ; Visser, R.G.F. ; Lee, T.A.J. van der; Vossen, J.H. - \ 2019
    Molecular Plant-Microbe Interactions 32 (2019)11. - ISSN 0894-0282 - p. 1536 - 1546.
    avirulence factors - cell death - chytridiomycota - effector - fungus-plant interactions - Genomics - hypersensitive response - metabolomics - plant-pathogen interactions - population genetics - proteomics - resistance genes
    Synchytrium endobioticum is an obligate biotrophic fungus of the phylum Chytridiomycota. It causes potato wart disease, has a world-wide quarantine status and is included on the HHS and USDA Select Agent list. S. endobioticum isolates are grouped in pathotypes based on their ability to evade host-resistance in a set of differential potato varieties. So far, thirty-nine pathotypes are reported. A single dominant gene (Sen1) governs pathotype 1 resistance and we anticipated that the underlying molecular model would involve a pathogen effector (AvrSen1) that is recognized by the host. The S. endobioticum specific secretome of fourteen isolates representing six different pathotypes was screened for effectors specifically present in pathotype 1(D1) isolates but absent in others. We identified a single AvrSen1 candidate. Expression of this candidate in potato Sen1 plants showed a specific hypersensitive response, which co-segregated with the Sen1 resistance in potato populations. No HR was obtained with truncated genes found in pathotypes that evaded recognition by Sen1. These findings established that our candidate gene was indeed Avrsen1. The S. endobioticum AvrSen1 is a single copy gene and encodes a 376 amino acid protein without predicted function or functional domains, and is the first effector gene identified in Chytridiomycota, an extremely diverse yet underrepresented basal lineage of fungi.
    The Solanum demissumR8 late blight resistance gene is an Sw-5 homologue that has been deployed worldwide in late blight resistant varieties
    Vossen, Jack H. ; Arkel, Gert van; Bergervoet-van Deelen, Marjan ; Jo, Kwang Ryong ; Jacobsen, Evert ; Visser, Richard G.F. - \ 2016
    Theoretical and Applied Genetics 129 (2016)9. - ISSN 0040-5752 - p. 1785 - 1796.
    Cisgenesis - Disease resistance gene - NB-LRR - Phytophthora infestans - Potato late blight

    The potato late blight resistance geneR8has been cloned.R8is found in five late blight resistant varieties deployed in three different continents. R8 recognises Avr8 and is homologous to the NB-LRR protein Sw-5 from tomato.Abstract: The broad spectrum late blight resistance gene R8 from Solanum demissum was cloned based on a previously published coarse map position on the lower arm of chromosome IX. Fine mapping in a recombinant population and bacterial artificial chromosome (BAC) library screening resulted in a BAC contig spanning 170 kb of the R8 haplotype. Sequencing revealed a cluster of at least ten R gene analogues (RGAs). The seven RGAs in the genetic window were subcloned for complementation analysis. Only one RGA provided late blight resistance and caused recognition of Avr8. From these results, it was concluded that the newly cloned resistance gene was indeed R8. R8 encodes a typical intracellular immune receptor with an N-terminal coiled coil, a central nucleotide binding site and 13 C-terminal leucine rich repeats. Phylogenetic analysis of a set of representative Solanaceae R proteins shows that R8 resides in a clearly distinct clade together with the Sw-5 tospovirus R protein from tomato. It was found that the R8 gene is present in late blight resistant potato varieties from Europe (Sarpo Mira), USA (Jacqueline Lee, Missaukee) and China (PB-06, S-60). Indeed, when tested under field conditions, R8 transgenic potato plants showed broad spectrum resistance to the current late blight population in the Netherlands, similar to Sarpo Mira.

    An updated conventional- and a novel GM potato late blight R gene differential set for virulence monitoring of Phytophthora infestans
    Suxian Zhu, Suxian ; Vossen, J.H. ; Bergervoet-van Deelen, J.E.M. ; Nijenhuis, M.A. ; Kodde, L.P. ; Kessel, G.J.T. ; Vleeshouwers, V.G.A.A. ; Visser, R.G.F. ; Jacobsen, E. - \ 2015
    Euphytica 202 (2015)2. - ISSN 0014-2336 - p. 219 - 234.
    resistance genes - solanum-bulbocastanum - disease resistance - durable resistance - rxlr effectors - united-states - pathogen - population - races - diversity
    Late blight is an important disease in potato that is caused by the oomycete Phytophthora infestans. In the past, Solanum demissum late blight resistance (R) genes were introgressed into cultivated potato (Solanum tuberosum). Eleven of these resistant plants were selected to characterize the virulence spectrum of individual P. infestans isolates and to monitor the dynamics of virulence in P. infestans populations. These plants are referred to as the Mastenbroek and Black differential sets. It has long been assumed that each differential plant contained one single R gene. In the current study and previous studies, however, most Mastenbroek differential plants were shown to harbor multiple R gene(s), which blurs virulence typing of late blight isolates. In order to acquire more accurate virulence profiles, we extended the Mastenbroek differential set with Solanum spp. plants harboring reduced R gene complexity and with plants containing recently identified R genes from related but different Solanum species. In addition, a differential set of ten Genetically Modified (GM) plants harboring single late blight R genes in the same genetic background (Desiree). By analyzing the virulence spectra of recently collected isolates using both newly described differential sets, we found that the GM Desiree differential set was more accurate for isolate virulence typing than the conventional (extended) differential set. Besides, the GM Desiree differential set was shown to be useful as trap plants to isolate novel P. infestans strains and to monitor virulence towards particular R genes in P. infestans populations `on site´. Legislative restrictions are, however, limiting the use of the GM Desiree differential set.
    Development of late blight resistant potatoes by cisgenic stacking
    Jo, K.R. ; Kim, C.J. ; Kim, S.J. ; Kim, T.J. ; Bergervoet-van Deelen, J.E.M. ; Jongsma, M.A. ; Visser, R.G.F. ; Jacobsen, E. ; Vossen, J.H. - \ 2014
    BMC Biotechnology 14 (2014). - ISSN 1472-6750
    broad-spectrum resistance - cultivar sarpo mira - phytophthora-infestans - solanum-bulbocastanum - r-gene - plants - transformation - genomics - tomato - biotechnology
    Background Phytophthora infestans, causing late blight in potato, remains one of the most devastating pathogens in potato production and late blight resistance is a top priority in potato breeding. The introduction of multiple resistance (R) genes with different spectra from crossable species into potato varieties is required. Cisgenesis is a promising approach that introduces native genes from the crops own gene pool using GM technology, thereby retaining favourable characteristics of established varieties. Results We pursued a cisgenesis approach to introduce two broad spectrum potato late blight R genes, Rpi-sto1 and Rpi-vnt1.1 from the crossable species Solanum stoloniferum and Solanum venturii, respectively, into three different potato varieties. First, single R gene-containing transgenic plants were produced for all varieties to be used as references for the resistance levels and spectra to be expected in the respective genetic backgrounds. Next, a construct containing both cisgenic late blight R genes (Rpi-vnt1.1 and Rpi-sto1), but lacking the bacterial kanamycin resistance selection marker (NPTII) was transformed to the three selected potato varieties using Agrobacterium-mediated transformation. Gene transfer events were selected by PCR among regenerated shoots. Through further analyses involving morphological evaluations in the greenhouse, responsiveness to Avr genes and late blight resistance in detached leaf assays, the selection was narrowed down to eight independent events. These cisgenic events were selected because they showed broad spectrum late blight resistance due to the activity of both introduced R genes. The marker-free transformation was compared to kanamycin resistance assisted transformation in terms of T-DNA and vector backbone integration frequency. Also, differences in regeneration time and genotype dependency were evaluated. Conclusions We developed a marker-free transformation pipeline to select potato plants functionally expressing a stack of late blight R genes. Marker-free transformation is less genotype dependent and less prone to vector backbone integration as compared to marker-assisted transformation. Thereby, this study provides an important tool for the successful deployment of R genes in agriculture and contributes to the production of potentially durable late blight resistant potatoes.
    Cloning and Characterization of R3b; Members of the R3 Superfamily of Late Blight Resistance Genes Show Sequence and Functional Divergence
    Li, G.C. ; Huang, S.W. ; Guo, X. ; Li, Y. ; Yang, Y. ; Guo, Z. ; Kuang, H.H. ; Rietman, H. ; Bergervoet-van Deelen, J.E.M. ; Vleeshouwers, V.V.G.A. ; Vossen, E.A.G. van der; Qu, D.Y. ; Visser, R.G.F. ; Jacobsen, E. ; Vossen, J.H. - \ 2011
    Molecular Plant-Microbe Interactions 24 (2011)10. - ISSN 0894-0282 - p. 1132 - 1142.
    broad-spectrum resistance - potato solanum-tuberosum - race-specific resistance - phytophthora-infestans - disease resistance - confers resistance - cultivated potato - plant - locus - bulbocastanum
    Massive resistance (R) gene stacking is considered to be one of the most promising approaches to provide durable resistance to potato late blight for both conventional and genetically modified breeding strategies. The R3 complex locus on chromosome XI in potato is an example of natural R gene stacking, because it contains two closely linked R genes (R3a and R3b) with distinct resistance specificities to Phytophthora infestans. Here, we report about the positional cloning of R3b. Both transient and stable transformations of susceptible tobacco and potato plants showed that R3b conferred full resistance to incompatible P infestans isolates. R3b encodes a coiled-coil nucleotide-binding site leucine-rich repeat protein and exhibits 82% nucleotide identity with R3a located in the same R3 cluster. The R3b gene specifically recognizes Avr3b, a newly identified avirulence factor from P. infestans. R3b does not recognize Avr3a, the corresponding avirulence gene for R3a, showing that, despite their high sequence similarity, R3b and R3a have clearly distinct recognition specificities. In addition to the Rpi-mcd1/Rpi-blb3 locus on chromosome IV, the R3 locus on chromosome XI is the second example of an R-gene cluster with multiple genes recognizing different races of P. infestans.
    Post-transciptional gene silencing of GBSS1 in potato: effects of size and sequence of the inverted repeats
    Heilersig, H.J.B. ; Loonen, A.E.H.M. ; Bergervoet-van Deelen, J.E.M. ; Wolters, A.M.A. ; Visser, R.G.F. - \ 2006
    Plant Molecular Biology 60 (2006)5. - ISSN 0167-4412 - p. 647 - 662.
    bound starch synthase - influences sirna efficacy - double-stranded-rna - solanum-tuberosum-l - transgenic plants - messenger-rna - virus-resistance - expression - granule - interference
    In the past, silencing of granule-bound starch synthase (GBSSI) in potato was achieved by antisense technology, where it was observed that inclusion of the 3¿ end of the GBSSI coding region increased silencing efficiency. Since higher silencing efficiencies were desired, GBSSI inverted repeat constructs were designed and tested in potato. First, large inverted repeats comprising the 5¿ and the 3¿ half of the GBSSI cDNA were tested. The 5¿ IR construct gave a significantly higher silencing efficiency than the 3¿ IR construct. Since it was not known whether the observed difference was due to the sequence or the orientation of the inverted repeat, the GBSSI cDNA was divided into three regions, after which each region was tested in small inverted repeats in two orientations. To this end large numbers of independent transformants were produced for each construct. The results suggested that there was no effect of inverted repeat orientation on silencing efficiency. The percentage of transformants showing strong inhibition varied from 48% for a 3¿-derived construct to 87% for a 5¿ as well as a middle region-derived construct. Similar to the large inverted repeats, the 3¿ sequences induced the least efficient silencing implying that the observed differences in silencing efficiency are caused by sequence differences. The small inverted repeat constructs with a repeat size of 500¿600 bp and a spacer of about 150 bp were more efficient silencing inducers than the large inverted repeat constructs where the size of the repeat was 1.1 or 1.3 kb whilst the size of spacer was 1.3 or 1.1 kb. The results presented here show that size and sequence of the inverted repeat influenced silencing efficiency.
    Isolation of protoplasts and culture and regeneration into plants in Alstroemeria
    Kim, J.B. ; Bergervoet-van Deelen, J.E.M. ; Raemakers, C.J.J.M. ; Jacobsen, E. ; Visser, R.G.F. - \ 2005
    In Vitro Cellular and Developmental Biology. Plant 41 (2005)4. - ISSN 1054-5476 - p. 505 - 510.
    somaclonal variation - cell-cultures - growth - callus - rice
    An efficient system for the regeneration of plants from protoplasts was developed in Alstroemeria. Friable embryogenic callus (FEC) proved to be the best source for protoplast isolation and culture when compared with leaf tissue and compact embryogenic callus. Protoplast isolation was most efficient when FEC was cultured under vacuum for 5 min in an enzyme solution consisting of 4% cellulase, 0.5% Driselase and 0.2% Macerozyme, followed by culture for 12¿16 h in the dark at 24°C. Cell wall formation and colony formation were better in a liquid medium than on a semi-solid agarose medium. Micro-calluses were formed after 4 wk of culture. Ninety percent of the micro-calluses developed into FEC after 12 wk of culture on proliferation medium. FEC cultures produced somatic embryos on a regeneration medium and half of these somatic embryos developed shoots. Protoplast-derived plants showed more somaclonal variation than vegetatively propagated control plants
    Regeneration of Pea (Pisum sativum L.) by a cyclic organogenic system
    Tzitzikas, E. ; Bergervoet-van Deelen, J.E.M. ; Raemakers, C.J.J.M. ; Vincken, J.P. ; Lammeren, A.A.M. van; Visser, R.G.F. - \ 2004
    Plant Cell Reports 23 (2004)7. - ISSN 0721-7714 - p. 453 - 461.
    somatic embryogenesis - transformation - thidiazuron - cultures - embryos - growth
    In a five-step procedure, plants were regenerated from meristematic tissue initiated from nodal tissue in four pea cultivars ('Espace', 'Classic', 'Solara', and 'Puget'). In step 1, stem tissue with one node (1-cm size) was subcultured on medium containing thidiazuron. As a result multiple shoots were produced, appearing normal or swollen at their bases. The multiple shoots were subcultured in the same medium, resulting in the formation of a green hyperhydric tissue in the swollen bases of the multiple shoots, which is fully covered with small buds [bud-containing tissue (BCT)]. In step 2, BCT fragments were isolated and subcultured in the same medium and, as a result, they were able to reproduce themselves in a cyclic fashion. In step 3, subculture of BCT on medium supplemented with a combination of gibberelic acid, 6-benzyladenine and a-naphthalene acetic acid (NAA), resulted in the formation of shoots, which were rooted in step 4 on medium supplemented with 0.5 mg/l NAA, indole-3-acetic acid (IAA) or indole-3-butyric acid. In step 5, in vitro plants were transferred to the greenhouse for acclimatisation and further development. The four varieties tested were all able to produce meristematic tissue, suggesting that its production is genotype independent.
    Isolation of a gene encoding a copper chaperone for the Cu/Zn superoxide dismutase and characterization of its promoter in Solanum tuberosum L.
    Trindade, L.M. ; Horvath, B.M. ; Bergervoet-van Deelen, J.E.M. ; Visser, R.G.F. - \ 2003
    Plant Physiology 133 (2003). - ISSN 0032-0889 - p. 618 - 629.
    solanum-tuberosum-l - oxidative stress - ascorbate peroxidase - antioxidant enzymes - regulated gene - abscisic-acid - water-stress - atx1 gene - expression - tuberization
    Gene expression during the potato (Solanum tuberosum) tuber lifecycle was monitored by cDNA-amplified fragment-length polymorphism, and several differentially expressed transcript-derived fragments were isolated. One fragment, named TDFL431, showed high homology to a copper (Cu) chaperone for Cu/zinc superoxide dismutase (CCS). The Ccs protein is responsible for the delivery of Cu to the Cu/zinc superoxide dismutase enzyme. The potato CCS (StCCS) full-length gene was isolated, and its sequence was compared with CCSs from other species. The promoter region of this gene was isolated, fused to the firefly luciferase coding sequence, and used for transformation of potato plants. The highest level of StCCS-luciferase expression was detected in the cortex of stem (like) tissues, such as stem nodes, stolons, and tubers; lower levels were detected in roots and flowers. The StCCS promoter contains regions highly homologous to several plant cis-acting elements. Three of them are related to auxin response, whereas four others are related to response to various stresses. Induction of the StCCS promoter was analyzed on 18 media, differing in hormone, sugar, and Cu content. StCCS expression was induced by auxin, gibberellins (GA(4 + 7)), fructose, sucrose, and glucose and was inhibited by relatively high concentrations of Cu.
    Exploring the use of cDNA-AFLP with leaf protoplasts as a tool to study primary cell wall biosynthesis in potato
    Oomen, R.J.F.J. ; Bergervoet-van Deelen, J.E.M. ; Bachem, C.W.B. ; Visser, R.G.F. ; Vincken, J.P. - \ 2003
    Plant Physiology and Biochemistry 41 (2003). - ISSN 0981-9428 - p. 965 - 971.
    carrot protoplasts - gene-expression - fusion
    An RNA fingerprinting study of potato leaf protoplasts was performed to explore its suitability for identifying candidate genes involved in primary cell wall biosynthesis. Microscopic analysis, using calcofluor white to stain cellulose, showed that the protoplasts generated a new cell wall in the first 18 h after transfer to a culture medium. Complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) was used to visualise differential gene expression at five distinct time-points within these first 18 h. In vitro plants (with and without exposure to severe physical damage) served as controls. Around 8500 transcript derived fragments (TDFs) were visualised which showed varying expression patterns in the protoplasts and controls. In total 156 TDFs were isolated, sequenced and used to search for homologies. Over 50% of these TDFs showed homology to described genes, involved in several general plant processes. However, only one cell wall related TDF (a pectin esterase) was found. Our results showed that even though the protoplasts actively regenerate a new cell wall, this did not result in highly increased expression of genes involved in cell wall biosynthesis or modification. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.
    Induction of recessive mutations in potato using tissue culture techniques
    Enckevort, L.J.G. van; Hoogkamp, T.J.H. ; Bergervoet-van Deelen, J.E.M. ; Visser, R.G.F. ; Jacobsen, E. ; Stiekema, W.J. ; Pereira, A. - \ 2001
    In: Joint FAO/IAEA Division of Nucelar Techniques in Food and Agriculture : In vitro techniques for selection of radiation induced mutations adapted to adverse environmental conditions, Shanghai, China, 1998 / International Atomic Energy Agency Wenen : IAEA - p. 15 - 25.
    Selection of independent Ds transposon insertions in somatic tissue of potato by protoplast regeneration
    Enckevort, L.J.G. van; Bergervoet-van Deelen, J.E.M. ; Stiekema, W.J. ; Pereira, A. ; Jacobsen, E. - \ 2000
    Theoretical and Applied Genetics 101 (2000). - ISSN 0040-5752 - p. 503 - 510.
    Potato is an autotetraploid crop plant that is not very amenable to the deployment of transposon tagging for gene cloning and gene identification. After diploidisation it is possible to get potato genotypes that grow well, but they are self-incompatible. This prevents the production of selfed progeny that are normally used in gene tagging approaches to select for parental lines with the target gene to be tagged in a homozygous stage. We describe here an alternative selection method for directed transposon tagging for a gene of interest in a heterozygous background. Diploid potato plants with a Ds transposon linked to the desired gene of interest (the Phytophthora infestans R1 resistance locus) in a heterozygous stage were used for the development of this directed transposon tagging strategy. After crossing to a diploid Ac transposon-containing genotype, 22 ’interesting' seedlings (R1Ds/r-; Ac/-) were selected that showed active Ds transposition as displayed by DNA blot hybridisation, empty donor site PCR and sequencing. Protoplast isolation and the use of the hygromycin gene as a cell-specific selection marker of Ds excision enabled the direct selection of Ds excision sectors in these highly chimaeric seedlings. This somatic selection of Ds transpositions and the regeneration through protoplasts resulted in the development of a large population of almost 2000 hygromycin-resistant plants. Southern blot analysis confirmed the insertion of Ds at independent positions in the genome. Every selected plant displayed independent Ds excisions and re-insertions due to the expression of the Ac transposase throughout development. This population, which is developed from seedlings with the desired R1 gene in a heterozygous stage, is directly useful for searching for transposon-tagged R1 mutants. In general, this approach for selecting for somatic transpositions is particularly suitable for the molecular isolation of genes in a heterozygous crop like potato.
    Genomic in situ hybridization identifies chromosome composition in the somatic hybrid Solanum tuberosum (+) Solanum nigrum and it's sexual and fusion progenies.
    Gavrilenko, T.A. ; Horsman, K. ; Bergervoet-van Deelen, J.E.M. ; Jacobsen, E. - \ 1998
    In: Proc. Intern. Symp. Breeding Res. on Potatoes, GroB Lusewitz (Rostock), Germany, Beitrage zur Zuchtungs-forschung 4 (1998) 49-5-0
    Plant regeneration from protoplasts isolated from friable embryogenic callus of cassava.
    Sofiari, E. ; Raemakers, C.J.J.M. ; Bergervoet-van Deelen, J.E.M. ; Jacobsen, E. ; Visser, R.G.F. - \ 1998
    Plant Cell Reports 18 (1998). - ISSN 0721-7714 - p. 159 - 165.
    Genomic in situ hybridization identifies chromosome composition in the somatic hybrid Solanum tuberosum (+) Solanum nigrum and it's sexual and fusion progenies.
    Gavrilenko, T.A. ; Horsman, K. ; Bergervoet-van Deelen, J.E.M. ; Jacobsen, E. - \ 1998
    In: Proceedings International Symposium Breeding Research on Potatoes, Gross Lusewitz, Rostock, Germany. Beitrage zur Zuchtungsforschung 4 - p. 49 - 50.
    Somatic hybridization between Solanum tuberosum and species of the Solanum nigrum complex: Selection of vigorously growing and flowering plants.
    Horsman, K. ; Bergervoet-van Deelen, J.E.M. ; Jacobsen, E. - \ 1997
    Euphytica 96 (1997). - ISSN 0014-2336 - p. 345 - 345.
    Methods for producing and transforming cassave protoplasts
    Visser, R.G.F. ; Raemakers, C.J.J.M. ; Jacobsen, E. ; Bergervoet-van Deelen, J.H.M. - \ 1996
    Octrooinummer: WO9744473, gepubliceerd: 1996.
    Method for producing protoplasts of cassave or closely related species, which protoplasts are capable of regeneration into plants. The method comprises producing friable embryogenic callus from explants of cassave of closely related species and isolating protoplasts from said friable embryogenic callus. Protoplasts which are obtainable by such a method. Method for transforming such a protoplast of cassave or closely related species, and transformed protoplast obtainable thereby. Method for regenerating plants from these protoplasts, and cassave plant or closely related species obtainable thereby.
    The first and second backcross progeny of the intergeneric fusion hybrids of potato and tomato after crossing with potato.
    Jacobsen, E. ; Daniel, M.K. ; Bergervoet-van Deelen, J.E.M. ; Huigen, D.J. ; Ramanna, M.S. - \ 1994
    Theoretical and Applied Genetics 88 (1994)2. - ISSN 0040-5752 - p. 181 - 186.
    Expression of a wild-type GBSS gene introduced into an amylose-free potato mutant by Agrobacterium tumefaciens and the inheritance of the inserts at the microsporic level.
    Flipse, E. ; Huisman, J.G.H. ; Vries, B.J. de; Bergervoet-van Deelen, J.E.M. ; Jacobsen, E. ; Visser, R.G.F. - \ 1994
    Theoretical and Applied Genetics 88 (1994). - ISSN 0040-5752 - p. 369 - 375.
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